Changes in Estimating the Wild Boar Carcasses Sampling Effort: Applying the EFSA ASF Exit Strategy by Means of the WBC-Counter Tool.

African swine fever (ASF) is a devastating disease, resulting in the high mortality of domestic and wild pigs, spreading quickly around the world. Ensuring the prevention and early detection of the disease is even more crucial given the absence of licensed vaccines. As suggested by the European Commission, those countries which intend to provide evidence of freedom need to speed up passive surveillance of their wild boar populations. If this kind of surveillance is well-regulated in domestic pig farms, the country-specific activities to be put in place for wild populations need to be set based on wild boar density, hunting bags, the environment, and financial resources. Following the indications of the official EFSA opinion 2021, a practical interpretation of the strategy was implemented based on the failure probabilities of wrongly declaring the freedom of an area even if the disease is still present but undetected. This work aimed at providing a valid, applicative example of an exit strategy based on two different approaches: the first uses the wild boar density to estimate the number of carcasses need to complete the exit strategy, while the second estimates it from the number of wild boar hunted and tested. A practical free access tool, named WBC-Counter, was developed to automatically calculate the number of needed carcasses. The practical example was developed using the ASF data from Sardinia (Italian island). Sardinia is ASF endemic from 43 years, but the last ASFV detection dates back to 2019. The island is under consideration for ASF eradication declaration. The subsequent results provide a practical example for other countries in approaching the EFSA exit strategy in the best choices for its on-field application.

Standardized Methodology for Target Surveillance against African Swine Fever.

African swine fever (ASF) remains the most serious pig infectious disease, and its persistence in domestic pigs and wild boar (WB) is a threat for the global industry. The surveillance of WB plays a central role in controlling the disease and rapidly detecting new cases. As we are close to eradicating ASF, the need to find any possible pockets of infection is even more important. In this context, passive surveillance is the method of choice for effective surveillance in WB. Considering the time and economic resources related to passive surveillance, to prioritize these activities, we developed a standardized methodology able to identify areas where WB surveillance should be focused on. Using GIS-technology, we divided a specific Sardinian infected area into 1 km2 grids (a total of 3953 grids). Variables related to WB density, ASF cases during the last three years, sex and age of animals, and the type of land were associated with each grid. Epidemiological models were used to identify the areas with both a lack of information and an high risk of hidden ASFV persistence. The results led to the creation of a graphic tool providing specific indications about areas where surveillance should be a priority.

Evaluation of the cost-effectiveness of ASF detection with or without the use of on-field tests in different scenarios, in Sardinia.

African swine fever (ASF) is a highly contagious disease of domestic pigs and wild boars (WBs). Without a vaccine, early antibody and antigen detection and rapid diagnosis are crucial for the effective prevention of the disease and the employment of control measures. In Sardinia, where 3 different suid populations coexisted closely for a long time, the disease persists since 1978. The recent ASF eradication plan involves more stringent measures to combat free-ranging pigs and any kind of illegality in the pig industry. However, critical issues such as the low level of hunter cooperation with veterinary services and the time required for ASF detection in the WBs killed during the hunting season still remain. Considering the need to deliver true ASF negative carcasses as early as possible, this study focuses on the evaluation and validation of a duplex pen-side test that simultaneously detects antibodies and antigens specific to ASF virus, to improve molecular diagnosis under field conditions. The main goal was to establish the specificity of the two pen-side tests performed simultaneously and to determine their ability to detect the true ASF negative carcasses among the hunted WBs. Blood and organ samples of the WBs hunted during the 2018/2019 hunting seasons were obtained. A total of 160 animals were tested using the pen-side kit test; samples were collected for virological and serological analyses. A specificity of 98% was observed considering the official laboratory tests as gold standards. The new diagnostic techniques could facilitate faster and cost-effective control of the disease.

Surveillance and control of African Swine Fever in free-ranging pigs in Sardinia.

African swine fever (ASF) is a notifiable infectious disease, caused by the ASF virus (ASFV), which is a DNA virus belonging to the family Asfarviridae, genus Asfivirus. This disease has gained importance in the last decade after its spread in several countries in Eastern and Central Europe, and more recently, in China. Despite the efforts made to eradicate it, ASF is still present on the Mediterranean island of Sardinia (Italy) and has been since 1978. ASF risk factors on the island have been analysed in previous studies; the role of free-ranging pigs in virus persistence has been suggested, but has not been fully elucidated. The most recent eradication plan provides more stringent measures to combat free-ranging pigs and any kind of illegality in the pig sector. From December 2017 to June 2018, a total of 29 depopulation actions were performed in 13 municipalities in central Sardinia, during which 2,281 free-ranging pigs were culled and more than 50% of them were tested for ASFV and antibody presence (1,218 and 1,416, respectively). A total of 651 pigs were seropositive, with a mean seroprevalence of 53.4% (CI 95% = 50.6-56.3), and 38 were ASFV positive (virus prevalence = 2.6%; CI 95% = 2.1-3.0). To the best of our knowledge, the present study is the first to provide a complete evaluation of this millennial system of pig farming and ASFV prevalence in free-ranging pigs. Furthermore, it has emphasised the necessity of combining the maintenance of an epidemiological surveillance program with continuous education of farmers and other people involved in pig husbandry, based on cultural and economic aspects.

[Monitoring plan of environmental contaminants in sheep milk from areas next to an Italian contaminated site named «Aree industriali di Porto Torres e discarica di Calancoi¼ (Industrial areas of Porto Torres and landfill of Calancoi) in Sardinia Region (Southern Italy)]. / Piano di monitoraggio per la ricerca di contaminanti ambientali in latte ovino prodotto in aree prossime al sito di interesse nazionale denominato «Aree industriali di Porto Torres e discarica di Calancoi¼ in Sardegna.

OBJECTIVES: to assess concentration levels of dioxins, PCBs, and chemical elements in sheep milk produced in areas close to the Italian contaminated site (CS) located in the North-West of Sardinia (Southern Italy), named «Aree industriali di Porto Torres e discarica di Calancoi¼ (i.e., industrial areas of Porto Torres and landfill of Calancoi). DESIGN: monitoring plan of environmental contaminants on zootechnical productions based on analysis of dioxins, PCBs, heavy metals (Cd, Hg, Pb), and other elements (As, Be, Cr, Ni, Tl , Zn) in sheep bulk milk. SETTING AND PARTICIPANTS: 60 samples of sheep bulk milk were collected in rural farms where animals were fed mainly with forage and feed produced in the CS: 30 samples from farms located near the industrial area of Porto Torres and 30 taken from farms spread out the landfill area of Calancoi, a former waste dump in the East of Sassari city. MAIN OUTCOME MEASURES: the concentration levels of target substances were determined and compared to legal thresholds fixed on EU regulations. RESULTS: concentrations of target substances in all samples were below the maximum limits set by EU regulations. Dioxins and DL-PCBs (expressed as the sum) were in the range 0.20-1.30 pg WHO-TEQ/g fat, NDL-PCB (sum of the six marker congeners) were in the range 0.50-7.50 ng/g, while concentrations of all heavy metals were not detectable or were near the limit of quantitation of the method. Undetectable concentrations were also observed for the toxic elements beryllium and thallium. CONCLUSIONS: concentration levels of micro-pollutants in milk samples were significantly lower than the legal thresholds. Concentrations and congener profiles showed no relevant differences compared to milk samples from areas with low environmental impact, therefore consumption of milk from the studied area do not produce a higher intake of contaminants respect to the regional milk. The results of this study do not make the resident population safe from health risks caused by critical issue of the CS, but provide information limited to the safety of the milk from these areas, without showing peculiar contamination and hazard for food from local sheep farms.